Allele-specific Diagnostic PCR Authentication of D.hancockii from the Other Dendrobium Species
LUO Yu ming 1, DING Xiao yu 2, XU Luo shan 3 BAO Shu lin 2 CHANG Jun 2 (1. Department of Biology, Huaiyin Teachers College, Huaian 223001, China) (2. College of Life Sciences, Nanjing Normal University, Nanjing 210097, China) (3. Department
Based on rDNA ITS sequences of D. hancockii and the other 37 species of Dendrobium, which are all used for the materials of various "Fengdou" and "Huangcao", the new allele specific diagnostic primers XYJB01S and XYJB01X have been designed to authenticate D. hancockii from the other species. Before the diagnostic PCR, the primer pair P1 and P2 for amplifying the whole ITS region should be used to validate template DNA at first so as to obtain the appropriate template DNA concentration for the diagnostic PCR. When the annealing temperature was raised to 64℃, only the template DNA of D. hancockii could be amplified whereas the diagnostic PCR of the other 37 species were all negative. The diagnostic PCR have been repeated for many times and have played an important role in identifying D. hancockii in China. Thus, the allele specific diagnostic primers have been designed to authenticate D. hancockii efficiently. Compared with the authentication method by sequencing DNA fragments, the allele specific diagnostic PCR was not only simple and timesaving but also practical and effective.